| Objective:To investigate the expression level of BRMS1L(breast cancer metastasis suppressor 1 like,BRMS1-like)in normal ovarian tissue and epithelial ovarian tumor and the correlation between BRMS1 L expression level and the clinicopathological features of epithelial ovarian cancer.After overexpression of BRMS1 L gene and inhibition of BRMS1 L gene expression by lentiviral transfection,serous adenocarcinoma cell line HeyA8 and ovarian mucinous adenocarcinoma cell line EFO27 biological and molecular biological characteristics changed,we could investigate the relationship between BRMS1 L and EMT in ovarian cancer,and provide a new direction to research for ovarian cancer metastasis.Methods:Immunohistochemistry was used to detect the expression level of BRMS1 L in 15 specimens of normal ovarian tissue and 65 specimens of epithelial ovarian tumor,and then analyse the relationship between BRMS1 L expression level and clinicopathological parameters of ovarian cancer.After transfection of BRMS1 L,Wound healing assay,Transwell migration and invasion assay,CCK8 assay,Plate clonality assay and Cell adhesion assay,Real-time fluorescence quantitative PCR,Western blot and Immunofluorescence assay were used to detect the expression level of BRMS1 L in mesothelial cell line HeyA8 and epithelial cell lines EFO27 and IGROV1 respectively,detect the changes of biological characteristics(proliferation,migration,invasion and adhesion)of ovarian cancer cell lines and explore the correlation between BRMS1 L and EMT in ovarian cancer,and at the same time investigate whether BRMS1 L via classical WNT signaling pathway to regulate EMT,thereby affecting the migration and invasion ability of ovarian cancer cells.Results:1.Compared with normal ovarian tissues,benign ovarian tumors and borderline ovarian tumors,the expression level of BRMS1 L in ovarian cancer was low and the difference was statistically significant(P<0.05).There was no significant correlation between the expression level of BRMS1 L in ovarian cancer group of age and histological type,but it was correlated with clinical stage,pathological grade and lymph node metastasis.(P<0.05).2.Compared with HeyA8(mesothelial cell morphology),the expression of BRMS1 L in EFO27 and IGROV1(epithelial cell morphology)was up-regulated with statistical significance(P <0.05).3.Wound healing assay,Transwell migration and invasion assay showed that the overexpression of BRMS1 L gene significantly inhibited the migration and invasion of ovarian cancer cell line HeyA8(P <0.01),inhibited the expression of BRMS1 L gene,The migration and invasion ability of ovarian cancer cell line EFO27 was significantly enhanced(P <0.01).4.The results of CCK8 assay,Plate clone assay and Cell adhesion assay showed that the proliferation and adhesive function of ovarian cancer cell line HeyA8 or EFO27 were not significantly different between overexpression of BRMS1 L gene and inhibition of BRMS1 L gene expression.5.After transfection of BRMS1 L,although EMT related markers vimentin(mesothelial cell marker)had no significant difference in protein expression level(mRNA expression level was significant),E-cadherin(epithelial cell marker)was significantly different in mRNA and protein expression level.6.After transfection of BRMS1 L gene,there was no significant difference in the expression level of ?-catenin and the mRNA expression level of TCF3/LEF1 in the classical WNT signaling pathway(WNT/?-catenin).Conclusions:1.The expression level of BRMS1 L in the tissues of the patients with ovarian cancer was lower than that of normal ovarian tissues,benign ovarian tumors and borderline ovarian tumors.The expression level of BRMS1 L in patients with ovarian cancer was not related with age,tumor histological type,but was related with clinical stage,pathological grade and lymph node metastasis,suggesting that the expression level of BRMS1 L in ovarian cancer tissues could reflect the degree of disease progression.2.The expression level of BRMS1 L in mesothelial cell line of ovarian cancer is lower than that of epithelial cell line.3.The migration and invasion ability of ovarian cancer cells were significantly inhibited with overexpression of BRMS1 L,while inhibiting the expression of BRMS1 L,the migration and invasion ability of ovarian cancer cells were significantly enhanced.However,that all had no significant effects on the proliferation ability and adhesion function of ovarian cancer cells,indicating that BRMS1 L gene was likely to be a tumor metastasis suppressor gene(BRMS1L only affected the migration and invasion ability of ovarian cancer cells but did not affect the proliferation ability).4.BRMS1 L may be associated with the EMT process of ovarian cancer and may be a potential target for the treatment of ovarian cancer and the improvement of prognosis.5.BRMS1 L did not regulate EMT via the classical WNT signaling pathway(WNT/?-catenin)to affect the migration and invasion of ovarian cancer cells. |
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